Schistosoma mansoni: relationship between cercarial production levels and snail host susceptibility

Two populations of Biomphalaria glabrata snails differing slightly in their susceptibility to Schistosoma mansoni infection showed dramatic differences in cercarial output per snail. Exposed to five or more miracidia, snails from a group with a 90-100% susceptibility rate (Group A) produced nearly twice the number of cercariae as those from a group with a 70-80% susceptibility rate (Group B). Exposure of individual snails to known numbers of miracidia resulted in higher numbers of primary (mother) sporocysts in Group A snails than in Group B snails. However, monomiracidial exposure of snails from both groups resulted in equivalent numbers of cercariae produced per positive snail, indicating that, once established, all primary sporocysts possess a similar reproductive potential. Morphometric analysis of serially sectioned 9-day-old primary sporocysts supported this conclusion; the size of the primary sporocysts and the size and numbers of secondary (daughter) sporocysts within each primary sporocyst were comparable in snails from both groups. The data indicate cercarial production in this system is regulated prior to, and/or during, early development of the primary sporocyst.     

A potential vector of Schistosoma mansoni in Uruguay

Susceptibility experiments were carried out with a Biomphalaria straminea-like planorbid snail (Biomphalaria aff. straminea, species inquirenda) from Espinillar, near Salto (Uruguay), in the area of the Salto Grande reservoir, exposed individually to 5 miracidia of Schistosoma mansoni (SJ2 and BH2 strains). Of 130 snails exposed to the SJ2 strain, originally infective to Biomphalaria tenagophila, 30 became infected (23%). The prepatent (precercarial) period ranged from 35 to 65 days. The cercarial output was irregular, following no definite pattern, varying from 138 to 76,075 per snail (daily average 4.3 to 447.5) and ending up with death. Three specimens that died, without having shed cercariae, on days 69 (2) and 80 after exposure to miracidia, had developing secondary sporocysts in their tissues, justifying the prospect of a longer precercarial period in these cases. In a control group of 120 B. tenagophila, exposed to the SJ2 strain, 40 became infected, showing an infection rate (33.3%) not significantly different from that of the Espinillar snail (chi 2 = 3.26). No cercariae were produced by any of the Espinillar snails exposed to miracidia of the BH2 strain, originally infective to Biomphalaria glabrata. Four specimens showed each a primary sporocyst in one tentacle, which disappeared between 15 and 25 days post-exposure, and two others died with immature, very slender sporocysts in their tissues on days 36 and 54. In a control group of 100 B. glabrata exposed to BH2 miracidia, 94 shed cercariae (94%) and 6 remained negative. Calculation of Frandsen's (1979a, b) TCP/100 index shows that "Espinillar Biomphalaria-SJ2 S. mansoni" is a vector-parasite "compatible" combination.(ABSTRACT TRUNCATED AT 250 WORDS)     

Schistosomatium douthitti: Exposure of Lymnaea catascopium to irradiated miracidia

Irradiation of Schistosomatium douthitti miracidia (4000, 5000, or 6000 rad) did not substantially alter their behavior or ability to penetrate their snail host. Treatment with 4000 rad was not sufficient to prevent all miracidia from establishing patent infections in Lymnaea catascopium, although significantly fewer snails exposed to these miracidia shed cercariae than did controls exposed to normal miracidia. Irradiation of miracidia with either 5000 or 6000 rad totally prevented cercarial production. Although destruction of irradiated mother sporocysts by encapsulating amebocytes was occasionally observed, most expanded without concomitant multiplication of germinal cells and embryo production and then collapsed. They generally persisted in this state throughout the period of observation (32 days). Snails sensitized by exposure to irradiated miracidia and challenged 2 or 10 days later with normal miracidia were as likely to develop patent infections as were snails exposed only to normal miracidia. Double sensitization of snails with irradiated miracidia also failed to confer protection upon challenge with normal miracidia. Most challenge sporocysts developed normally, often in close proximity to collapsed irradiated sporocysts.     

Larval development of Fasciola hepatica in experimental infections: variations with populations of Lymnaea truncatula

A retrospective study was undertaken on 70 French populations of Lymnaea truncatula experimentally infected with Fasciola hepatica to determine whether or not susceptibility of snails to infection influenced redial and cercarial production. Results were compared with those obtained from two control populations, known for prevalences higher than 60% when experimentally infected with F. hepatica. In the 70 other populations examined, the prevalences ranged from 2 to 75%. In 55 of these populations, where the prevalence was more than 20%, a high proportion (50.1-56.8%) of snails died after cercarial shedding, whereas in the other groups (non-shedding snails with the most differentiated larvae being free cercariae, rediae containing cercariae, immature rediae, or sporocysts, respectively), snail death was significantly less. In 11 populations, where the prevalence values were 5-19%, only 14% of snails died after cercarial shedding, whereas snails with free cercariae, rediae with cercariae, or immature rediae showed significant increases in snail mortality. In the remaining four snail populations, with prevalences of less than 5%, the most differentiated larval forms were only immature rediae and/or sporocysts. Overall, the number of rediae containing cercariae significantly decreased with decreasing prevalence values. The low prevalence of experimental infection in several populations of snails might be explained by the occurrence of natural infections with miracidia originating from a mammalian host other than cattle, and/or by genetic variability in the susceptibility of snails to infection.     

Transplantation of Schistosoma japonicum daughter sporocysts in Oncomelania hupensis

Schistosoma japonicum daughter sporocysts obtained from infected Oncomelania hupensis hupensis were successfully transplanted to parasite-free O. hupensis hupensis. Survival and infection rates of recipient snails were 80% and 75% respectively. Intramolluscan development of transplanted daughter sporocysts in recipient snails appears to proceed in a similar manner as those reported for transplanted S. mansoni and S. haematobium in their respective snail intermediate hosts. Complete colonization of the digestive gland of recipient snails by sporocysts was observed 80 days after transplantation. Cercarial production during a 10-day observation from recipient snails was characterized by periods of high and low and irregular daily emissions. The average daily cercarial production was 150 per snail. Cercariae produced by recipient snails were infective to mice. Of those cercariae exposed to mice, approximately 30% developed to adult schistosomes. These results have definitive utility in the maintenance of S. japonicum in the laboratory.     

Schistosoma mansoni: characterization of clones maintained by the microsurgical transplantation of sporocysts

Five male and 5 female clones of Schistosoma mansoni were established and maintained for 3 yr by the serial microsurgical transplantation of sporocysts from infected to uninfected Biomphalaria glabrata snails. The clones were initially derived from 10 randomly selected snails with monomiracidial infections. Clones were characterized by several criteria, including their infectivities for mice and snails, their cercarial outputs, and their ability to produce immunity in mice. The mean infectivities of individual clones in mice ranged from 26 to 44%, and were highly consistent within each clone. The infectivities of cloned sporocysts in snails ranged from 44 to 100% and were also highly consistent within clones. Mean cercarial outputs from individual clones ranged from 450 to 4,300 per snail. In mice, clones differed significantly from each other in their ability to immunize and in their susceptibility to immunity. Each clone was unique and did not appear to differ with time or subpassaging through snails, suggesting that the differences had a genetic basis.     

Schistosoma mansoni infections in neonatal Biomphalaria glabrata snails.

In areas endemic for schistosomiasis, the population dynamics of the snail intermediate hosts have a direct effect on parasite transmission. The present study focused on the potential for neonatal Biomphalaria glabrata snails to become infected with Schistosoma mansoni and to produce cercariae under various conditions. It was found that snails as small as 0.74 mm in shell diameter could survive miracidial penetration and could release cercariae when as small as 1.6 mm in diameter. Cercariae produced by small snails were equally infectious for mice when compared with those shed by larger snails. Likewise, histological examination of neonatally exposed snails revealed normally developing parasites at all stages of infection. It was found that in 2 snail populations expressing either high or low susceptibility to the parasite, peak susceptibility occurred at 25 days of age in both groups. Daily cercarial production for neonatally exposed snails was initially low but increased dramatically as the snails grew, eventually reaching values as high as 2,100 cercariae/snail/day. A moderate to high percentage of snails infected as neonates was eventually capable of simultaneously producing both eggs and cercariae. These studies emphasize the potential importance of neonatal and preadult snails in helping to maintain foci of S. mansoni infection in endemic areas.     

The effect of size of M line Biomphalaria glabrata on the course of development of Echinostoma paraensei

M line Biomphalaria glabrata snails of 4-, 6-, 8-, 10-, 12-, or 20-mm shell diameter were individually exposed to 10 miracidia each of Echinostoma paraensei. Snails 10 mm in size or larger were found to be significantly less likely to harbor intraventricular sporocysts than snails in smaller size categories. The percentage of snails with intraventricular sporocysts that also developed hemocyte encapsulation responses generally increased with snail size, whereas the number of snails that ultimately became heavily parasitized with large numbers of daughter rediae decreased significantly with snail size. However, at least some snails in each size category developed such disseminated infections. Comparative histological study of 6- and 12-mm snails revealed that parasites readily penetrated both groups of snails, but were more likely to be encapsulated and destroyed in larger snails. Encapsulation reactions were noted from 1 to 15 days postexposure (dpe) in 12-mm snails, indicating that unlike other commonly studied models of trematode-gastropod interactions, snail resistance is not always manifested during the first few days following exposure. Upon infection with E. paraensei, both 6- and 12-mm snails showed significant increases in the number of circulating hemocytes/mm3 of hemolymph. In 6-mm snails, such increases occurred concurrently with successful parasite development. Hemocyte counts in 6-mm snails were significantly elevated from 4 to 15 dpe whereas in 12-mm snails they were significantly elevated from 2 to 30 dpe. A significant degree of resistance to E. paraensei develops as B. glabrata grows and attains sexual maturity. A mechanistic understanding of this phenomenon awaits further investigation.     

Selection of genotypes of Schistosoma mansoni and their maintenance by sporocyst transplantation

Schistosoma mansoni was isolated by hatching eggs obtained from a naturally infected Rat in Grand Etang, Guadeloupe; fifty Biomphalaria glabrata were exposed to five miracidia each. The resulting cercariae were used to infect laboratory mice which were later sacrificed to provide worms for enzyme analyses and eggs for further infections. Seven enzymes in extracts of individual worms were examined by isoelectric focusing in polyacrylamide gels: AcP, G6PDH, PGM, GPI and HK showed no variation, whereas MDH and LDH proved to be polymorphic. Two MDH loci were recognised, MDH-2 was invariant whereas two alleles were assumed at the MDH-1 locus. It was not possible to make a genetic interpretation of the complex banding pattern of LDH, although 4 types (LDH-A, -B, -C, -D) were observed. Of the snail infections, one batch of snails was exposed to 5 miracidia per snail in the normal way whereas other snails were each exposed to a single miracidium. The latter were sacrificed to provide sporocysts to transplant into further groups of recipient snails. Cercariae from the recipient snails were used to infect mice and the adult worms were analysed and compared with the normally passaged material. In this way, three lines, defined by the possession of particular MDH and LDH types, were selected from the originally polymorphic population; two were identical. The combination of single miracidium infections and enzyme typing has illustrated the possibility of selecting parasite lines of known genotype; transplantation of sporocysts from snail to snail has demonstrated that such lines can be maintained exclusively in the intermediate host.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of water temperature on the ability of Diplostomum spathaceum miracidia to establish in lymnaeid snails

The effects of water temperature on the ability of Diplostomum spathaceum miracida to infect and establish patent infections in Lymnaea peregra and L. stagnalis were investigated. Snails were infected over a range of temperatures (6-20 degrees C) and kept thereafter at 20 degrees C or were infected at 20 degrees C and kept at either 14, 20, or 25 degrees C. Infection success was determined after 8 weeks by either observing cercarial shedding or examining snail viscera for sporocysts. The establishment of miracidia declined at lower water temperatures despite maintenance for 8 weeks at 20 degrees C while exposure of snails to miracidia at 20 degrees C and maintenance at different temperatures had little apparent effect. Infection success under these conditions was related more to the numbers of miracidia to which the snails were exposed. However, under this latter experimental regime, the time taken for the infection to become patent clearly depended upon maintenance temperature.     

Schistosomatium douthitti: effects of Lymnaea catascopium age on susceptibility to infection

Laboratory-reared Lymnaea catascopium snails (1–269 days old) were exposed individually to different numbers of Schistosomatium douthitti miracidia. Increasing the exposure dosage from 3 to 10 miracidia generally increased infection rates, in some age classes up to 100%. Successful re-exposure of snails not infected after a primary exposure was possible. Neonatal snails were least likely to become infected, primarily because miracidia were not attracted to them. Snails 12–55 days old were most susceptible to infection. Miracidia were readily attracted to these snails, and many were ingested and subsequently penetrated the host esophageal wall. Miracidial penetration of external snail surfaces was rare. Susceptibility of older snails (65–269 days) progressively declined with age. Many miracidia were entangled and immobilized in mucus produced by these snails, and fewer were ingested. No conspicuous host cellular responses to mother sporocysts were observed in any of the snails sectioned. A comparison of susceptibility of deliberately stunted snails and comparably aged controls of normal size indicated that the former were more susceptible.     

Sublethal effects of ultraviolet b radiation on miracidia and sporocysts of Schistosoma mansoni: intramolluscan development, infectivity, and photoreactivation

Schistosoma mansoni occurs in tropical regions where levels of ultraviolet B (UVB; 290-320 nm) light are elevated. However, the effects of UVB on parasite transmission are unknown. This study examines effects of UVB on the miracidia and sporocysts of S. mansoni, focusing specifically on intramolluscan development, infectivity, and the ability to photoreactivate (repair DNA damage using visible light). Histology revealed that miracidia irradiated with 861 J x m(-2) underwent abnormal development after penetrating Biomphalaria glabrata snails. Total number of sporocysts in snail tissues decreased as a function of time postinfection (PI), among both nonirradiated and irradiated parasites; however, this decrease was greater in the latter. Moreover, whereas the proportion alive of nonirradiated sporocysts increased PI, that of irradiated sporocysts, i.e., derived from irradiated miracidia, decreased. Irradiation of miracidia with UVB resulted in decreased prevalence of patent infection (defined by presence of daughter sporocysts) in a dose-dependent manner, and no infections occurred at a dose of 861 J x m(-2). Like many aquatic organisms, including the snail host, parasites were able to photoreactivate if exposed to visible light following UVB irradiation, even subsequent to penetrating snails. These photoreactivation results suggest cyclobutane-pyrimidine dimers in DNA as the primary mechanism of UVB damage, and implicate photoreactivation, rather than nucleotide excision, as the main repair process in S. mansoni.     

Schistosoma mansoni: effect of infection on reproduction and gonadal growth in Biomphalaria glabrata

Sexually mature Biomphalaria glabrata were exposed to 12 miracidia of Schistosoma mansoni, and egg production of snails was monitored over a period of 5 weeks. During the study period, exposed snails grew at approximately the same rate as unexposed controls. Castration, as measured by a reduction in the mean number of eggs laid per snail occurred between 14 and 21 days postexposure (PE). The reduction in fecundity in infected snails coincided with the migration and establishment of daughter sporocysts in the digestive gland and gonad. Enumeration of individual oocytes in longitudinal sections of the ovotestis revealed that uninfected snails contained significantly more oocytes per section than infected snails at 27, 31, and 40 days PE. In addition, the mean area of gonadal sections of control snails increased over the 40-day experimental period, whereas there was no such increase in gonadal area of infected snails. These data suggest that there is an inhibition in gonadal growth in infected snails. When oocyte data were expressed in terms of mean gonadal area, the mean number of oocytes per mm2 of gonad of uninfected and infected snails did not differ significantly over the study period, except at Day 14 PE, when infected snails contained a significantly greater number of oocytes per mm2 of gonad than did uninfected controls. It is hypothesized that daughter sporocysts of S. mansoni are primarily responsible for the inhibition of host reproductive activity, and may be mediating their effects through mechanisms involved in the regulation of gonadal growth.(ABSTRACT TRUNCATED AT 250 WORDS)     

Cercarial shedding of Echinostoma cinetorchis and experimental infection of the cercariae to several kinds of snails

The development of Echinostoma cinetorchis in several snail species reared in laboratory aquaria was observed. The eggs from adult flukes collected from the intestine of rats were cultivated to miracidia, and exposed to Hippeutis sp. snails. Observations were made for cercarial shedding from the exposed snails. The cercariae shed from the snails were again exposed to several species of fresh water snails in order to observe metacercarial formation in the snails and their infectivity to final hosts. The results obtained in this study were as follows: 1. Twenty miracidia were exposed to each snail of Hippeutis sp. About 58.3% of the above snails (7 out of 12) were dead before shedding the cercariae, and the remainder shed the cercariae for a period of 7 to 9 days before death. 2. Cercarial shedding from the infected snails started from the 25th day after the exposure to miracidia, and the total number of cercariae shed per snail was 684 in average (range; 482-904). 3. The size of rediae developed in the infected Hippeutis sp. snails was 1,242 x 214 microns in average, and the number of rediae per snail was 350 in average (range; 120-510). 4. About 40 to 50 cercariae shed from the Hippeutis sp. snails were each exposed to several species of snails reared in the laboratory. The metacercarial formation was confirmed by dissecting the infected snails, 12 to 16 days after the infection.(ABSTRACT TRUNCATED AT 250 WORDS)     

Fasciola hepatica: characteristics of infection in Lymnaea truncatula in relation to the number of miracidia at exposure.

Experimental infections of Lymnaea truncatula by Fasciola hepatica were carried out in three snail populations to determine whether the number of miracidia used for each snail at exposure (1, 2, 5, 10, or 20 per snail) had any influence on the characteristics of Fasciola infection and metacercarial production. The number of miracidia had a significant influence on snail survival at day 30 postexposure and the frequency of infected L. truncatula that died without shedding (NCS snails). The frequency of NCS snails, the growth of cercaria-shedding snails throughout the experiment, the time between exposure and the first cercarial shedding, the duration of shedding, and the number of metacercariae were independent of the number of miracidia used for each snail. The highest metacercaria productivity for each miracidium was found in single-miracidium infections. Single-miracidium infections were the most effective, as the mean number of cercariae was the same as in other groups, whereas their survival rate was much higher.     

Schistosoma mansoni: male-biased sex ratios in snails and mice

Adult sex ratios of Schistosoma mansoni, in mice, were shown to be biased toward males (3:1) despite the finding that sex ratios of miracidia were 50:50. The adult male bias was caused by greater male infectivity of miracidia for snails and cercariae for mice. A significantly higher percentage of male miracidia developed to cercarial production in unimiracidial infections (57 male, 34 female), and a significantly higher percentage of male cercariae developed to adulthood in mice (143 male, 79 female worms resulted from 900 male and 900 female cercariae). No significant differences were found between male and female parasites for longevity of miracidia (both sexes, 10 hr) and cercariae (males 21.3 +/- 5.75 hr, females 25.0 +/- 7.02 hr), prepatent periods in snail hosts (male 34 +/- 2.92 days, females 33 +/- 2.36 days), longevity of snail infections (males 96.6 +/- 25.15 days, females 115.2 +/- 82.43 days), and the numbers of cercariae produced per snail lifetime (males 30,751.44 +/- 18,064.33, females 34,083.00 +/- 33,732.82). Present results provide a better understanding of the life cycle of S. mansoni, are of theoretical significance for theories of biased sex ratios (which at present cannot account for the male-biased ratio of S. mansoni), and also suggest that schistosomiasis transmission models assuming a 50:50 sex ratio at all stages of the life cycle should be reassessed.     

The invasion of Biomphalaria pfeifferi by Schistosoma mansoni miracidia and the development of daughter sporocysts

Laboratory experiments have been carried out to determine the susceptibility of Gezira Biomphalaria pfeifferi snails to S. mansoni miracidia and the relationship between miracidia and daughter sporocyst production at the 10–17 day development stage. The relationship between snail numbers, miracidia numbers and water volume has also been studied. Two non susceptible snails, Bulinus truncatus and Cleopatra bulimoides, both of which occur naturally in Gezira canals, were tested to see if they act as decoys for S. mansoni miracidia.The results showed that the B. pfeifferi are 100% susceptible to S. mansoni invasion, at least to the daughter sporocyst development stage. The more miracidia that penetrated the more daughter sporocysts were produced, however individual variation and overlap were great. When one miracidium was released to find one snail it succeeded in low water volumes (5 m, 50 ml), but failed in 5 litres. When 100 miracidia were released mortality of snails was high suggesting superinfection particularly when only one or five snails were available. Among survivors daughter sporocyst counts were very high. Cleopatra and Bulinus snails do have a decoy effect when present in large numbers. In their presence the number of infected snails was marginally reduced and the number of daughter sporocysts greatly reduced. However, if superinfection is reduced by decoy effect, it is conceivable that Biomphalaria may be protected by decoy snails in circumstances where miracidia counts are high.     

Pathology and host responses induced by Schistosomatium douthitti in the freshwater snail Lymnaea catascopium.

Most Schitosomatium douthitti miracidia penetrated the esophageal wall of Lymnaea catascopium without provoking amoebocyte encapsulation responses or extensive pathological changes. Amoebocytes frequently attached to developing mother and daughter sporocysts, but did not encapsulate or destroy them. Pressure resulting from extensive growth of mother sporocysts ruptured the transverse membrane of some snails. After releasing daughter sporocysts, mother sporocysts in some snails were destroyed by amoebocytes. Many migrating cercariae became trapped in the tissues of L. catascopium, particularly in the posterior portion of the foot, and were encapsulated and destroyed. Large increases in numbers of amoebocytes in the anterior portion of the lung roof of infected snails were noted, even before cercarial production had been initiated. Atrophy of the digestive gland occurred in infected snails.     

Influence of shell size of Lymnaea columella on infectivity and development of Fasciola hepatica

Experimental infections of Lymnaea columella with Fasciola hepatica were carried out to determine the influence of shell size on the infection rate and on the outcome of rediae and cercariae. Snails were divided into seven groups according to shell size: 2-4 mm, 5-6 mm, 7-8 mm, 9-10 mm, 11-12 mm, 13-14 mm and 15 mm or more. One hundred snails in each group were infected by using four miracidia for each snail. Snails with larger shell size showed a lower infection rate, the groups presenting the highest (79%) and lowest (2%) proportions of positives being those of 5-6 mm and 15 mm or more, respectively. Cercariae were present in 21% of them at 31 days post-infection, and cercarial shedding was observed 61 days post-infection. It was concluded that there is a non-linear negative association between shell size and infection rate.     

Schistosoma mansoni: Long-term maintenance of clones by microsurgical transplantation of sporocysts

Schistosoma mansoni sporocysts originally derived from monomiracidially infected Biomphalaria glabrata snails were serially transplanted into the cephalopedal sinus of anesthetized snails by the microsurgical implantation of fragments of parasitized hepato-pancreas and ovotestis. Three to six passages each of five male and five female clones were maintained for as long as 2.0 years. Of the recipient snails which survived surgery, 87% released cercariae, usually beginning 5–7 weeks after surgery. The percentage of snails which released cercariae increased with successive passages. The mean survival time of surgically infected snails after cercarial emergence began was 9.2 ± 0.5 weeks, nearly the same as that of miracidially infected snails. Longevities of snails infected with male or female clones were similar. Recipient snail size and age did not influence cloning success. Beginning 5 weeks from the onset of cercarial emergence large numbers of cercariae (a mean of 3900/snail from male clones and 1300/snail from females) were obtained during each shedding period. These results clearly demonstrate that the microsurgical transplantation of sporocysts is a practical means of maintaining and expanding populations of genetically homogeneous schistosomes (clones).